骨髓内皮细胞条件培养液促进小鼠胚胎干细胞生成造血集落形成细胞

目的:为观察骨髓内皮细胞务件培养液(mouse bone marrow stromal cell-conditional medium,mBMEC-CM)对鼠胚胎干细胞生成造血集落形成细胞的影响.方法:将鼠胚胎干细胞系D3细胞(embry-onic stem cell line-D3,ES-D3)形成4 d拟胚体(day-4 embryoid bodies,4dEBs),再用mBMEC-CM诱导4dEBs生成高增殖潜能集落形成细胞(high proliferation potential-colony formation cell,HPP-CFC)和红系爆式集落形成单位(burst forming unit-erythroid,BFU-E).以形成造血集落的数量为检测指标,观察mBMEC-CM诱导浓度、天数和诱导生成的细胞数与形成HPP-CFC和BFU-E数之间的关系.结果:形成的HPP-CFC和BFU-E集落数均与诱导生成的4dEBs细胞数呈正相关(HPP-CFC:r=0.916,P＜0.05;BFU-E:r=0.927,P＜0.05),且均随着种入的细胞数(在1×107～4×107/L范围内)增加而呈现相应的增加.当种入的细胞数增加到5×107/L时两种集落数均不再增加.mBMEC-CM诱导浓度(在0～20%范围内)与其诱导生成的HPP-CFC和BFU-E数呈剂量依赖性正相关(HPP-CFC:r=0.909,P＜0.05;BFU-E:r=0.927,P＜0.01).20%浓度mBMEC-CM诱导4dEBs来源的细胞于3,6和9 d形成的HPP-CFC和BFU-E数,以诱导3 d者最高,6 d次之,9 d最低.结论:骨髓内皮细胞条件培养液能促进鼠胚胎干细胞分化为HPP-CFC和BFU-E.

Bone marrow endothelial cell-conditional medium promotes the generation of hematopoietic colony-forming cells from murine embryonic stem cells

OBJECTIVE: To observe the inductive efficiency of deriving hematopoietic colony-forming cells from murine embryonic stem (mES) cells co-cultured with bone marrow stromal cell-conditional medium (mBMEC-CM). METHODS: After the day-4 embryoid bodies (4 dEBs) were derived from embryonic stem cell-D3 (ES-D3) cells, the cells of 4 dEBs were induced into hematopoietic colony-forming cells by co-culturing with mBMEC-CM. The numbers of 4 dEB-derived hematopoietic colonies (high proliferation potential-colony formation cells and burst forming unit-erythroid, HPP-CFC and BFU-E) were detected to explore the relation between the implanted 4 dEB-derived cell numbers and the colony numbers of BFU-E and HPP-CFC. The inducing effect of mBMEC-CM was observed according to the doses and days of induction. RESULTS: The number of 4 dEB-derived cells within 1 x 10(7)-4 x 10(7)/L was positively related to the colony numbers of HPP-CFC and BFU-E (HPP-CFC, r=0.916,P< 0.05; BFU-E, r=0.927, P<0.05). The inducing doses of mBMEC-CM within 0-20% were positively related to the colony numbers of HPP-CFC and BFU-E (HPP-CFC, r=0.909, P<0.05; BFU-E, r=0.927, P<0.01). The colony numbers of HPP-CFC and BFU-E derived from the 4 dEB-derived cells were the highest after 3 days of induction, followed by those of 6 days and 9 days. CONCLUSION: Bone marrow endothelial cell-conditional medium can promote the generation of HPP-CFC and BFU-E from murine embryonic stem cells.